A framework for assessing confidence in metabarcoding assays and results

PLEASE NOTE: The closing date for this notice has been extended to 18/09/2024. DNA-based methods have the potential to significantly change how we monitor and assess ecosystems. These methods are now being used more widely within Natural England, particularly the detection of ecological communities using environmental DNA (eDNA). This is often conducted by employing metabarcoding assays which target specific species assemblages (e.g. fish, mammals) or broader taxonomic groups (e.g. vertebrates, invertebrates, bacteria). It is difficult for Natural England staff to understand, interpret, and determine confidence in the results of different metabarcoding assays or assess the risk of false positive or false negative results. This project aims to address some of these challenges through the development of an eDNA metabarcoding assay validation framework. This will include: establishing key criteria to enable the classification of metabarcoding assays based on their taxonomic coverage, taxonomic resolution and detection sensitivity for different groups; and assessing the suitability of metabarcoding assays for different sample types and environments. This will enable end-users to determine the recommended scenarios for application of a given assay and improve assay performance with further validation.